Methods of inhibiting tumor metastasis

ABSTRACT

Thromboxane A2 receptor antagonists, such as ifetroban, inhibit solid tumor metastasis. The formation of surface and microscopic lung metastases are inhibited. Thromboxane A2 receptor antagonists can inhibit the tumor metastasis process without affecting the growth or development of a primary tumor.

RELATED APPLICATIONS

This patent application is a continuation of U.S. patent applicationSer. No. 17/941,371, filed Sep. 9, 2022, which is a continuation of U.S.patent application Ser. No. 16/767,534, filed May 27, 2020, which is theU.S. national stage entry, under 35 U.S.C. § 371, of InternationalApplication Number PCT/US2018/062967, filed Nov. 29, 2018, which claimspriority to U.S. Provisional Application No. 62/592,365, filed Nov. 29,2017, the entire contents of each of which are incorporated herein byreference.

TECHNICAL FIELD

The invention provides methods of inhibiting solid tumor metastasis withthromboxane A2 receptor antagonists.

BACKGROUND

Cancer remains a leading cause of morbidity and mortality globally. Forthe majority of cancers, cytotoxic chemotherapy has been the standard ofcare for decades, although emerging advances in targeted therapies andimmunotherapeutic strategies are altering the reliance on chemotherapyfor some cancers. Despite advances in how some cancers are treated,cancer metastasis and recurrence are universal concerns across all solidcancers diagnosed in early stage.

Metastatic cancer diagnoses impose a significant emotional, social, andeconomic burden that impacts quality of life for each patient, in largepart because these diagnoses generally associate with early death. Evenwhen a cancer is in remission (a clinically silent state), microscopicmetastases, even single cancer cells, can lurk clinically undetectablethroughout the body for variably long periods of time, depending on thetype of cancer. These potentially dangerous cells remain unseen, even tothe most sensitive imaging modalities, until they grow to a level thatbecomes clinically apparent.

Few therapies directly target tumor cell invasion and/or metastasis andnone are currently in FDA-approved clinical use for this indication.Deliberate approaches to specifically prevent metastases by neutralizingor thwarting the problematic dispersion of tumor cells would beimportant, and could manifest in a number of different forms:containment of cells to the primary site, disabling cancer cell survivalin the circulation, reducing evasion of immune surveillance, preventingplatelet/tumor cell aggregates, preventing extravasation at secondarysites, and/or making secondary sites inhospitable to metastatic seeding.Agents that specifically affect the metastatic process across cancerswould thus represent a new paradigm in cancer management, addressing anunmet medical need and aimed ultimately at blocking dissemination ofcancers—focusing beyond the primary tumor.

Blockade of metastasis may have additional indirect benefits as well.For example, agents that thwart the spread of cancer might allowconventional chemotherapies to be used at reduced doses or for shorterdurations. This is an important consideration, since chemotherapiesoften target fast-growing healthy cells in addition to fast growingtumor cells, causing treatment-associated toxicity, tissue damage, andmorbidity. Further, treatments aimed at preventing metastaticdissemination might be used in combination with molecularly targetedanti-tumor agents. In this scenario, it should be noted that acquiredresistance to molecularly targeted agents often arises at sites ofmetastatic recurrence. Thus, inhibition of metastatic dissemination mayreduce the incidence and/or rate of acquired resistance, improving theclinical success of precision cancer therapeutics.

Patients diagnosed with solid cancers might receive any one or acombination of treatments, including surgery, radiation, chemotherapy,vaccines, hormone modulators, immunotherapies, or molecularly targetedtherapies. After the treatment course, including the treatment of anymicrometastatic disease with adjuvant systemic therapy, a patient mayhave no remaining clinical evidence of cancer. In these cases, thecancer is considered to be ‘in remission’. These patients, theirfamilies, and their physicians rely on watchful waiting until the timeclinical evidence of cancer returns. Prevention of initial or secondarymetastatic spread of any remaining tumor cells—by targeting themetastatic process itself—could replace watchful waiting with proactiveprevention.

There is a need therefore for effective therapies that block or inhibitthe spread or metastasis of solid tumors from a primary tumor site.

SUMMARY

In one aspect, the invention provides a method of inhibiting solid tumormetastasis comprising administering to a subject in need thereof, anamount of a thromboxane A2 receptor antagonist effective to inhibitmetastasis of a solid tumor in the subject.

In another aspect, the invention provides a thromboxane A2 receptorantagonist, or a pharmaceutically acceptable salt or compositionthereof, for use in the treatment or inhibition of solid tumormetastasis in a subject.

In another aspect, the invention provides a thromboxane A2 receptorantagonist, or a pharmaceutically acceptable salt or compositionthereof, for use in a method of treating or inhibiting solid tumormetastasis, wherein the method comprises administering the thromboxaneA2 receptor antagonist to a subject in need thereof.

In another aspect, the invention provides a use of a thromboxane A2receptor antagonist, or a pharmaceutically acceptable salt orcomposition thereof, for the preparation of a medicament for thetreatment or inhibition of solid tumor metastasis in a subject.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the effects of ifetroban to decrease surface lungmetastases from orthotopic primary 4T1 tumors in mice.

FIG. 2 shows the effects of ifetroban to decrease microscopic lungmetastases from orthotopic primary 4T1 tumors in mice.

FIG. 3 shows the effects of ifetroban and aspirin to decrease metastasisin the 4T1 mouse model of metastasis.

FIG. 4 shows the effects of ifetroban to decrease lung metastases inMDA-MB-231 mice.

FIG. 5A shows that ifetroban has no effect on 4T1 cell growth andviability in vitro.

FIG. 5B shows that primary tumor volume in the 4T1 mouse model is notaffected by ifetroban.

FIG. 5C shows that total mouse body weight is not affected by ifetroban.

FIG. 6 shows a conceptual rendering of a proposed solid tumor metastasisprocess.

FIG. 7 shows primary tumor sites that resulted in the metastatic spreadin patients having the gain of function variant T399A of the thromboxaneA2 receptor.

FIG. 8 shows decreased hematogenous metastasis of breast, pancreatic andlung cancer cells following administration of ifetroban.

DETAILED DESCRIPTION

Cancer Metastasis

Cancers can spread at different rates and employ distinct molecular andcellular pathways in one patient as compared to others. Some of thisvariability may be attributable to a host environment that is more orless conducive to metastatic spread. In contrast to local tissueinvasion by cancer cells, metastasis requires the transport of cancercells to distant sites via the blood and/or lymphatic system; perineuralinvasion can also play an important role in a number of cancers. Cancermetastasis may involve the detachment and embolization of tumor cellaggregates, which may be increased in size via interaction withhematopoietic cells within the circulation. Other aspects of metastasisinclude the circulation of tumor emboli within the vasculature (bothhematologic and lymphatic), the survival of tumor cells that traffickedthrough the circulation and arrest in a capillary bed, and theextravasation of the tumor embolus, by mechanisms similar to thoseinvolved in the initial tissue invasion.

“Metastasis of a tumor/cancer cell”, as used herein, refers to thedissemination/transmission of a tumor/cancer cell from an original siteto one or more noncontiguous sites elsewhere in the body, e.g., from oneorgan or part to another not directly connected with it by way of, forexample, blood vessels or lymphatics. The metastasis of a tumor/cancercell can, for example, lead to the formation of a secondary orsubsequent tumor at a site other than the location of the primary tumor.The tumor/cancer cell of the inventive methods can be a cell of anysolid tumor/cancer, such as those described herein.

Distinct from the metastasis process (i.e., dissemination/transmission)is the growth of a metastatic tumor/cancer at a secondary site, whichcan involve proliferation of the tumor cells within the organ parenchymaresulting in a metastatic focus, establishment of vascularization, anddefenses against host immune responses.

Selected platelet-mediated steps in metastasis that it may be possibleto target via pharmacological agents include: detachment andembolization of platelet-containing tumor cell aggregates; circulatingtumor cells protected from host immune responses by aggregatedplatelets; and tumor embolus colonization facilitated byP-selectin-mediated tumor cell-platelet interactions.

While a number of investigations indicate involvement of platelets inthe path to cancer metastasis (see FIG. 6 ), research exploring themanipulation of these connections to inhibit platelet activation withthe deliberate goal of preventing or interrupting metastatic diseaseremains at a very early stage. No such proven therapies yet exist. Thediverse ways in which platelets interact with tumor cells and thecomplex directionality of those effects are yet incompletely understoodand FIG. 6 is not intended to be an exhaustive representation of thisfield; for example, much remains unexplored regarding interactionsbetween platelets and tumor cells, the role that other factors may playin the overall disease process, and the potential benefit ofantiplatelet therapeutics, including factors such as intercellulartransfer of mitochondrial DNA and various observed epigenetic changes.

Thromboxane A2 receptor antagonists may have broad utility in solidtumor metastasis inhibition or prevention, across multiple tumor/cancertypes. Without being bound by a particular theory, thromboxane A2receptor antagonists decrease platelet aggregation and may decrease theability of tumor cells to detach from the vascular endothelium andattach to platelets. Through blockade of platelet aggregation with tumorcells, thromboxane A2 receptor antagonists may decrease circulatingtumor cell survival through decreased integrin- and/or selectin-mediatedcell survival signaling. Thromboxane A2 receptor antagonists may preventcolonization by affecting P-selectin-mediated interactions. Plateletsmay shield circulating tumor cell clusters from host immune responses.Through blockade of platelet aggregation with tumor cells, thromboxaneA2 receptor antagonists may disrupt the interactions between plateletsand tumor cell clusters, thereby increasing exposure of circulatingtumor cells to host immune responses and inhibiting metastasis.Thromboxane A2 receptor antagonists may inhibit the formation ofcirculating tumor cell clusters, the movement of circulating tumor cellclusters, and/or the aggregation of circulating tumor cell clusters withplatelets.

Methods of Inhibiting Tumor Metastasis

One aspect of the invention encompasses a method for inhibiting tumorcell metastasis in a subject. The method comprises administering anamount of a thromboxane A2 receptor antagonist effective to inhibitmetastasis of a solid tumor in the subject.

The invention also provides a thromboxane A2 receptor antagonist, or apharmaceutically acceptable salt or composition thereof, for use in thetreatment or inhibition of solid tumor metastasis in a subject. Theinvention also provides a thromboxane A2 receptor antagonist, or apharmaceutically acceptable salt or composition thereof, for use in amethod of treating or inhibiting solid tumor metastasis, wherein themethod comprises administering the thromboxane A2 receptor antagonist toa subject in need thereof. The treatment or inhibition of solid tumormetastasis comprises use of the thromboxane A2 receptor antagonist in anamount effective to inhibit metastasis of a solid tumor in a subject.

The invention also provides the use of a thromboxane A2 receptorantagonist, or a pharmaceutically acceptable salt or compositionthereof, for the preparation of a medicament in the treatment orinhibition of solid tumor metastasis in a subject. The invention alsoprovides the use of a thromboxane A2 receptor antagonist, or apharmaceutically acceptable salt or composition thereof, for thepreparation of a medicament in a method of treating or inhibiting solidtumor metastasis, wherein the method comprises administering thethromboxane A2 receptor antagonist to a subject in need thereof. Thetreatment or inhibition of solid tumor metastasis comprises use of thethromboxane A2 receptor antagonist in an amount effective to inhibitmetastasis of a solid tumor in a subject.

Inhibition of tumor cell metastasis may be manifested by a reduction inthe overall metastatic burden, e.g., the number, distribution, or volumeof metastatic tumors in the treated subject relative to an untreatedsubject. In one embodiment, the number of metastatic tumors may bereduced at least two-fold. In another embodiment, the number ofmetastatic tumors may be reduced at least ten-fold. In still anotherembodiment, the number of metastatic tumors may be reduced at least50-fold. In yet another embodiment, the number of metastatic tumors maybe reduced at least 200-fold. In a further embodiment, the number ofmetastatic tumors may be reduced to such an extent such that nometastatic tumors are detectable. In still another embodiment,metastatic tumors may be restricted to one organ or tissue, rather thanbeing spread to two or more organs or tissues.

Inhibition of metastasis may be measured by many parameters inaccordance with the present invention and, for instance, may be assessedby delayed appearance of secondary tumors, slowed development ofsecondary tumors, decreased occurrence of secondary tumors, slowed ordecreased severity of secondary effects of disease, arrested tumorgrowth, reduced rate of metastatic recurrence, and regression of tumors,among others. In the extreme, complete inhibition is referred to hereinas prevention. In addition, the inhibition of metastasis may beidentified by a reduction in metastatic foci present in the animal. Instill another embodiment, inhibition of metastasis may manifest as aperiod of metastasis free survival in a treated subject. In someembodiments, a thromboxane A2 receptor antagonist is administered in anamount effective to effective to reduce the rate of metastaticrecurrence.

In some embodiments, a thromboxane A2 receptor antagonist isadministered in an amount effective to inhibit metastasis of a solidtumor in a subject without inhibiting the growth/development of thesolid tumor itself, i.e., at a primary/secondary site. Thus, thethromboxane A2 receptor antagonist may inhibit the metastatic processwithout affecting the growth or development of a primary/secondarytumor.

The subject may be a mammal, such as a human patient. Non-human animalsinclude companion animals, such as cats, dogs and horses. Other types ofnon-human animals envisioned for treatment according to the presentmethods include commercially important animals, including sheep, swine,cattle and others.

The primary cancer may be a cancer of epithelial origin. The primarycancer type may be lung cancer, non-small cell lung cancer, breastcancer, ovarian cancer, prostate cancer, testicular cancer, pancreaticcancer, melanoma, sarcoma, cervical cancer, endometrial cancer, livercancer, uterine cancer, kidney (renal)cancer, gastroesophageal cancer,colon cancer, bladder cancer, mouth cancer, or throat cancer.

The types of metastasis that may be inhibited and/or eliminated includemetastasis to the lung and/or bone (such as to the spine). It isenvisioned that the present methods and preparations will also findutility in reducing and/or preventing the metastasis of tumor/cancercells to other organs, such as, by way of example and not limitation,metastasis to ovary, liver, brain, kidney, spleen, intestines, adrenalglands, or any other tissue and/or organ or combination of tissuesand/or organs.

Thromboxane A2 receptor antagonists suitable for use in the disclosedmethods include, but are not limited to ifetroban, GR32191, SQ29548,sulotroban, daltroban, linotroban, ramatroban, seratrodast, terutroban,Z-235, LCB-2853, SQ28668, ICI 192605, AH23848, CPI-211, or pinane TXA₂.Suitable thromboxane A2 receptor antagonists are also described in U.S.Pat. No. 5,100,889, which is incorporated herein by reference.

For example, the thromboxane A2 receptor antagonist may have formula (I)

-   -   including all stereoisomers thereof, wherein    -   m is 1, 2, or 3;    -   n is 0, 1, 2, 3, or 4;

-   -   Z is —(CH₂)₂—, —CH═CH—, or    -   Y is O, a single bond, or —CH═CH—;    -   R is CO₂H, CO₂C₁₋₆alkyl, CH₂OH, —CONHSO₂R³, —CONHR^(3a), or        —CH₂-tetrazol-5-yl;    -   R³ is C₁-6alkyl, 6- to 10-membered aryl, or L¹-(6- to        10-membered aryl);    -   R^(3a) is C₁₋₆alkyl, 6- to 10-membered aryl, or L¹-(6- to        10-membered aryl);    -   X is O, S, or NH;    -   R¹ is hydrogen, C₁₋₈alkyl, C₂₋₈alkenyl, C₂₋₈alkynyl,        —(CH₂)_(t)-C(O)—NHR^(1a), or —(CH₂)^(t)—NH—C(O)R^(1a);    -   G¹ is a 6- to 10-membered aryl, a C₃₋₁₂cycloalkyl, a 5- to        12-membered heteroaryl, or a 4- to 12-membered heterocyclyl;    -   L¹ is C₁₋₆alkylene;    -   t is 1 to 12;    -   R^(1a) is C₁₋₆alkyl, C₃₋₁₂cycloalkyl, 42-C₃₋₁₂cycloalkyl, or a        6- to 10-membered aryl;    -   R² is hydrogen, C₁₋₆alkyl, 6- to 10-membered aryl, or 42-(6- to        10-membered aryl);    -   or R¹ and R² together with the nitrogen to which they attach        form a 4- to 8-membered heterocyclic ring;    -   wherein each aryl is independently and optionally substituted        with 1 or 2 substituents independently selected from the group        consisting of halogen, cyano, C₁₋₆alkyl, C1-6haloalkyl, OH,        —OC₁₋₆alkyl, —OC1-6haloalkyl, —SC₁₋₆alkyl, —S(O)C₁₋₆alkyl,        —S(O)₂C₁₋₆alkyl, —OC₁₋₆ alkylene-phenyl, —S-phenyl,        —S(O)-phenyl, and —S(O)₂-phenyl;    -   wherein each cycloalkyl is independently and optionally        substituted with 1-4 substituents independently selected from        the group consisting of halogen, C₁₋₆alkyl, C₁₋₆haloalkyl, OH,        and —OC₁₋₆alkyl.

Thromboxane A2 receptor antagonists include compounds of formula (I-a),(I-b), (I-c), (I-d), (I-e), and (I-f), wherein Z¹ is —(CH₂)₂— or—CH═CH—.

In some embodiments Z¹ is —CH═CH—. In further embodiments, Z¹ is—CH═CH—, m is 1 and n is 2. In still further embodiments, Z¹ is —CH═CH—and R is CO₂H.

In formula (I), Z may be

In some embodiments, formula (I) has formula (I-g), wherein R, R¹, R²,X, m, and n are as defined herein.

In some embodiments, formula (I) has formula (I-h), wherein R, R¹, R²,X, m, and n are as defined herein.

Included in the formulas (I) and (I-a) to (I-h) are compounds wherein mis 1, n is 2, R is CO₂H, R² is hydrogen, and R¹ is C₁₋₈alkyl. Accordingto any compounds of formulas (I), (I-g), or (I-h) are compounds whereinX is O.

The thromboxane A2 receptor antagonist may be in the form of apharmaceutically acceptable salt. For example, in formulas (I) and (I-a)to (I-h), the group CO₂H at R may be in the form of an alkali metalsalt, such as the sodium salt (i.e., R=CO₂Na).

The compound may exist as a stereoisomer wherein asymmetric or chiralcenters are present. The stereoisomer is “R” or “S” depending on theconfiguration of substituents around the chiral carbon atom. The terms“R” and “S” used herein are configurations as defined in IUPAC 1974Recommendations for Section E, Fundamental Stereochemistry, in PureAppl. Chem., 1976, 45: 13-30. The disclosure contemplates variousstereoisomers and mixtures thereof and these are specifically includedwithin the scope of this invention. Stereoisomers include enantiomersand diastereomers, and mixtures of enantiomers or diastereomers.Individual stereoisomers of the compounds may be prepared syntheticallyfrom commercially available starting materials, which contain asymmetricor chiral centers or by preparation of racemic mixtures followed bymethods of resolution well-known to those of ordinary skill in the art.These methods of resolution are exemplified by (1) attachment of amixture of enantiomers to a chiral auxiliary, separation of theresulting mixture of diastereomers by recrystallization orchromatography and optional liberation of the optically pure productfrom the auxiliary as described in Furniss, Hannaford, Smith, andTatchell, “Vogel's Textbook of Practical Organic Chemistry,” 5th edition(1989), Longman Scientific & Technical, Essex CM20 2JE, England, or (2)direct separation of the mixture of optical enantiomers on chiralchromatographic columns, or (3) fractional recrystallization methods.

The present disclosure also includes an isotopically-labeled compound,which is identical to those recited in formula (I), but for the factthat one or more atoms are replaced by an atom having an atomic mass ormass number different from the atomic mass or mass number usually foundin nature. Examples of isotopes suitable for inclusion in the compoundsof the invention are hydrogen, carbon, nitrogen, oxygen, phosphorus,sulfur, fluorine, and chlorine, such as, but not limited to ²H, ³H, ¹³C,¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively.Substitution with heavier isotopes such as deuterium, i.e. ²H, canafford certain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements and, hence, may be preferred in some circumstances. Thecompound may incorporate positron-emitting isotopes for medical imagingand positron-emitting tomography (PET) studies for determining thedistribution of receptors. Suitable positron-emitting isotopes that canbe incorporated in compounds of formula (I) are ¹¹C, ¹³N, ¹⁵O, and ¹⁸F.Isotopically-labeled compounds of formula (I) can generally be preparedby conventional techniques known to those skilled in the art or byprocesses analogous to those described in the accompanying Examplesusing appropriate isotopically-labeled reagent in place ofnon-isotopically-labeled reagent.

The term “alkyl,” as used herein, means a straight or branched,saturated hydrocarbon chain. The term “lower alkyl” or “C₁₋₆alkyl” meansa straight or branched chain hydrocarbon containing from 1 to 6 carbonatoms. The term “C1-4alkyl” means a straight or branched chainhydrocarbon containing from 1 to 4 carbon atoms. Representative examplesof alkyl include, but are not limited to, methyl, ethyl, n-propyl,iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl,isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl,2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl.

The term “alkenyl,” as used herein, means a straight or branched,hydrocarbon chain containing at least one carbon-carbon double bond.

The term “alkylene,” as used herein, refers to a divalent group derivedfrom a straight or branched chain hydrocarbon, for example, of 1 to 6carbon atoms. Representative examples of alkylene include, but are notlimited to, —CH₂CH₂—, —CH₂CH₂CH₂—, —CH₂CH₂CH₂CH₂—, and—CH₂CH₂CH₂CH₂CH₂—.

The term “aryl,” as used herein, refers to a phenyl or a phenyl appendedto the parent molecular moiety and fused to a cycloalkyl group (e.g.,indanyl), a phenyl group (i.e., naphthyl), or a non-aromatic heterocycle(e.g., benzo[d][1,3]dioxol-5-yl).

The term “cycloalkyl,” as used herein, refers to a saturated ring systemcontaining all carbon atoms as ring members and zero double bonds. Acycloalkyl may be a monocyclic cycloalkyl (e.g., cyclopropyl), a fusedbicyclic cycloalkyl (e.g., decahydronaphthalenyl), or a bridgedcycloalkyl in which two non-adjacent atoms of a ring are linked by analkylene bridge of 1, 2, 3, or 4 carbon atoms (e.g.,bicyclo[2.2.1]heptanyl). Representative examples of cycloalkyl include,but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantyl,and bicyclo[1.1.1]pentanyl.

The term “halogen” or “halo,” as used herein, means Cl, Br, I, or F.

The term “haloalkyl,” as used herein, means an alkyl group, as definedherein, in which one, two, three, four, five, six, seven or eighthydrogen atoms are replaced by a halogen.

The term “heteroaryl,” as used herein, refers to an aromatic monocyclicheteroatom-containing ring (monocyclic heteroaryl) or a bicyclic ringsystem containing at least one monocyclic heteroaryl (bicyclicheteroaryl). The monocyclic heteroaryl are five or six membered ringscontaining at least one heteroatom independently selected from the groupconsisting of N, and S (e.g. 1, 2, 3, or 4 heteroatoms independentlyselected from O, S, and N). The five membered aromatic monocyclic ringshave two double bonds and the six membered six membered aromaticmonocyclic rings have three double bonds. The bicyclic heteroaryl is an8-to 12-membered ring system having a monocyclic heteroaryl ring fusedto a monocyclic aromatic or carbocyclic ring, a monocyclic heteroaryl,or a monocyclic heterocycle. The bicyclic heteroaryl is attached to theparent molecular moiety at an aromatic ring atom. Representativeexamples of heteroaryl include, but are not limited to, indolyl (e.g.,indol-1-yl, indol-2-yl, indol-4-yl), pyridinyl (including pyridin-2-yl,pyridin-3-yl, pyridin-4-yl), pyrimidinyl, pyrazinyl, pyridazinyl,pyrazolyl (e.g., pyrazol-4-yl), pyrrolyl, benzopyrazolyl,1,2,3-triazolyl (e.g., triazol-4-yl), 1,3,4-thiadiazolyl,1,2,4-thiadiazolyl, 1,3,4-oxadiazolyl, 1,2,4-oxadiazolyl, imidazolyl,thiazolyl (e.g., thiazol-4-yl), isothiazolyl, thienyl, benzimidazolyl(e.g., benzimidazol-5-yl), benzothiazolyl, benzoxazolyl,benzoxadiazolyl, benzothienyl, benzofuranyl, isobenzofuranyl, furanyl,oxazolyl, isoxazolyl, purinyl, isoindolyl, quinoxalinyl, indazolyl(e.g., indazol-4-yl, indazol-5-yl), quinazolinyl, 1,2,4-triazinyl,1,3,5-triazinyl, isoquinolinyl, quinolinyl,6,7-dihydro-1,3-benzothiazolyl, imidazo[1,2-c]pyridinyl (e.g.,imidazo[1,2-a]pyridin-6-yl), naphthyridinyl, pyridoimidazolyl,thiazolo[5,4-b]pyridin-2-yl, thiazolo[5,4-d]pyrimidin-2-yl.

The term “heterocycle” or “heterocyclic,” as used herein, means amonocyclic heterocycle, a bicyclic heterocycle, or a tricyclicheterocycle. The monocyclic heterocycle is a three-, four-, five-, six-,seven-, or eight-membered ring containing at least one heteroatomindependently selected from the group consisting of O, N, and S. Thethree- or four-membered ring contains zero or one double bond, and oneheteroatom selected from the group consisting of N, and S. Thefive-membered ring contains zero or one double bond and one, two orthree heteroatoms selected from the group consisting of O, N and S. Thesix-membered ring contains zero, one or two double bonds and one, two,or three heteroatoms selected from the group consisting of O, N, and S.The seven- and eight-membered rings contains zero, one, two, or threedouble bonds and one, two, or three heteroatoms selected from the groupconsisting of O, N, and S. Representative examples of monocyclicheterocycles include, but are not limited to, azetidinyl, azepanyl,aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl,1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl,isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl,2-oxo-3-piperidinyl, 2-oxoazepan-3-yl, oxadiazolinyl, oxadiazolidinyl,oxazolinyl, oxazolidinyl, oxetanyl, oxepanyl, oxocanyl, piperazinyl,piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl,pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl,tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, 1,2-thiazinanyl,1,3-thiazinanyl, thiazolinyl, thiazolidinyl, thiomorpholinyl,1,1-dioxidothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, andtrithianyl. The bicyclic heterocycle is a monocyclic heterocycle fusedto a phenyl group, or a monocyclic heterocycle fused to a monocycliccycloalkyl, or a monocyclic heterocycle fused to a monocycliccycloalkenyl, or a monocyclic heterocycle fused to a monocyclicheterocycle, a monocyclic heterocycle fused to a monocyclic heteroaryl,or a spiro heterocycle group, or a bridged monocyclic heterocycle ringsystem in which two non-adjacent atoms of the ring are linked by analkylene bridge of 1, 2, 3, or 4 carbon atoms, or an alkenylene bridgeof two, three, or four carbon atoms. The bicyclic heterocycle isattached to the parent molecular moiety at a non-aromatic ring atom(e.g., 2-oxaspiro[3.3]heptan-6-yl, indolin-1-yl,hexahydrocyclopenta[b]pyrrol-1(2H)-yl). Representative examples ofbicyclic heterocycles include, but are not limited to, benzopyranyl,benzothiopyranyl, chromanyl, 2,3-dihydrobenzofuranyl,2,3-dihydrobenzothienyl, 2,3-dihydroisoquinoline,2-azaspiro[3.3]heptan-2-yl, 2-oxa-6-azaspiro[3.3]heptan-6-yl,azabicyclo[2.2.1]heptyl (including 2-azabicyclo[2.2.1]hept-2-yl),azabicyclo[3.1.0]hexanyl (including 3-azabicyclo[3.1.0]hexan-3-yl),2,3-dihydro-1H-indolyl, isoindolinyl, octahydrocyclopenta[c]pyrrolyl,octahydropyrrolopyridinyl, and tetrahydroisoquinolinyl. Tricyclicheterocycles are exemplified by a bicyclic heterocycle fused to a phenylgroup, or a bicyclic heterocycle fused to a monocyclic cycloalkyl, or abicyclic heterocycle fused to a monocyclic cycloalkenyl, or a bicyclicheterocycle fused to a monocyclic heterocycle, or a bicyclic heterocyclein which two non-adjacent atoms of the bicyclic ring are linked by analkylene bridge of 1, 2, 3, or 4 carbon atoms, or an alkenylene bridgeof two, three, or four carbon atoms. Examples of tricyclic heterocyclesinclude, but are not limited to, octahydro-2,5-epoxypentalene,hexahydro-2H-2,5-methanocyclopenta[b]furan,hexahydro-1H-1,4-methanocyclopenta[c]furan, aza-adamantane(1-azatricyclo[3.3.1.13,7]decane), and oxa-adamantane(2-oxatricyclo[3.3.1.13,7]decane). The monocyclic, bicyclic, andtricyclic heterocycles are connected to the parent molecular moiety at anon-aromatic ring atom.

Terms such as “alkyl,” “cycloalkyl,” “alkylene,” etc. may be preceded bya designation indicating the number of atoms present in the group in aparticular instance (e.g., “C1-4alkyl,” “C3-6cycloalkyl,”“C1-4alkylene”). These designations are used as generally understood bythose skilled in the art. For example, the representation “C” followedby a subscripted number indicates the number of carbon atoms present inthe group that follows. Thus, “C3alkyl” is an alkyl group with threecarbon atoms (i.e., n-propyl, isopropyl). Where a range is given, as in“C₁₋₄,” the members of the group that follows may have any number ofcarbon atoms falling within the recited range. A “C1-4a1kyl,” forexample, is an alkyl group having from 1 to 4 carbon atoms, howeverarranged (i.e., straight chain or branched).

In some embodiments, the method of inhibiting solid tumor metastasisincludes administering an amount of a thromboxane A2 receptor antagonisteffective to inhibit metastasis in combination with one or morechemotherapeutic agents. Chemotherapeutic agent refers to a chemicalcompound that is useful in the treatment of cancer. The compound may bea cytotoxic agent that affects rapidly dividing cells in general, or itmay be a targeted therapeutic agent that affects the deregulatedproteins of cancer cells. The chemotherapeutic agent may be analkylating agent, an anti-metabolite, an anti-tumor antibiotic, ananti-cytoskeletal agent, a topoisomerase inhibitor, an anti-hormonalagent, a targeted therapeutic agent, an immunotherapy, or a combinationthereof. Non-limiting examples of alkylating agents include altretamine,benzodopa, busulfan, carboplatin, carboquone, carmustine, chlorambucil,chlomaphazine, cholophosphamide, chlorozotocin, cisplatin,cyclosphosphamide, dacarbazine (DTIC), estramustine, fotemustine,ifosfamide, improsulfan, lomustine, mechlorethamine, mechlorethamineoxide hydrochloride, melphalan, meturedopa, nimustine, novembichin,phenesterine, piposulfan, prednimustine, ranimustine; temozolomide,thiotepa, triethylenemelamine, trietylenephosphoramide,triethylenethiophosphaoramide, trimethylolomelamine, trofosfamide,uracil mustard and uredopa. Suitable anti-metabolites include, but arenot limited to aminopterin, ancitabine, azacitidine, 6-azauridine,capecitabine, carmofur, cytarabine or cytosine arabinoside (Ara-C),dideoxyuridine, denopterin, doxifluridine, enocitabine, floxuridine,fludarabine, 5-fluorouracil (5-FU), gemcetabine, leucovorin (folinicacid), 6-mercaptopurine, methotrexate, pemetrexed, pterop-terin,thiamiprine, trimetrexate, and thioguanine. Non-limiting examples ofsuitable anti-tumor antibiotics include aclacinomysin, actinomycin,adriamycin, authramycin, azaserine, bleomycins, cactinomycin,calicheamicin, carabicin, caminomycin, carzinophilin, chromomycins,dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine,doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin,mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin,potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin,streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin.Non-limiting examples of suitable anti-cytoskeletal agents includecolchicines, docetaxel, macromycin, paclitaxel (taxol), vinblastine,vincristine, vindesine, and vinorelbine. Suitable topoisomeraseinhibitors include, but are not limited to, amsacrine, etoposide(VP-16), irinotecan, RFS 2000, teniposide, and topotecan. Non-limitingexamples of suitable anti-hormonal agents such as aminoglutethimide,aromatase inhibiting 4(5)imidazoles, bicalutamide, finasteride,flutamide, goserelin, 4-hydroxytamoxifen, keoxifene, leuprolide, LY117018, mitotane, nilutamide, onapristone, raloxifene, tamoxifen,toremifene, and trilostane. No-limiting examples of targeted therapeuticagents include a monoclonal antibody such as alemtuzumab, bevacizumab,capecitabine, cetuximab, gemtuzumab, heregulin, rituximab, trastuzumab;a tyrosine kinase inhibitor such as imatinib mesylate; and a growthinhibitory polypeptide such as erythropoietin, interleukins (e.g., IL-1,IL-2, IL-3, IL-6), leukemia inhibitory factor, interferons,thrombopoietin, TNF-α, CD30 ligand, 4-1 BB ligand, and Apo-1 ligand.Also included are pharmaceutically acceptable salts, acids, orderivatives of any of the above listed agents. The mode ofadministration of the chemotherapeutic agent can and will vary dependingupon the agent and the type of tumor or neoplasm. A skilled practitionerwill be able to determine the appropriate dose of the chemotherapeuticagent.

The disclosed compounds may exist as pharmaceutically acceptable salts.The term “pharmaceutically acceptable salt” refers to salts orzwitterions of the compounds which are water or oil-soluble ordispersible, suitable for treatment of disorders without undue toxicity,irritation, and allergic response, commensurate with a reasonablebenefit/risk ratio and effective for their intended use. The salts maybe prepared during the final isolation and purification of the compoundsor separately by reacting an amino group of the compounds with asuitable acid. For example, a compound may be dissolved in a suitablesolvent, such as but not limited to methanol and water and treated withat least one equivalent of an acid, like hydrochloric acid. Theresulting salt may precipitate out and be isolated by filtration anddried under reduced pressure. Alternatively, the solvent and excess acidmay be removed under reduced pressure to provide a salt. Representativesalts include acetate, adipate, alginate, citrate, aspartate, benzoate,benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate,digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate,formate, isethionate, fumarate, lactate, maleate, methanesulfonate,naphthylenesulfonate, nicotinate, oxalate, pamoate, pectinate,persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate,propionate, succinate, tartrate, trichloroacetate, trifluoroacetate,glutamate, para-toluenesulfonate, undecanoate, hydrochloric,hydrobromic, sulfuric, phosphoric and the like. The amino groups of thecompounds may also be quaternized with alkyl chlorides, bromides andiodides such as methyl, ethyl, propyl, isopropyl, butyl, lauryl,myristyl, stearyl and the like.

Basic addition salts may be prepared during the final isolation andpurification of the disclosed compounds by reaction of a carboxyl groupwith a suitable base such as the hydroxide, carbonate, or bicarbonate ofa metal cation such as lithium, sodium, potassium, calcium, magnesium,or aluminum, or an organic primary, secondary, or tertiary amine.Quaternary amine salts can be prepared, such as those derived frommethylamine, dimethylamine, trimethylamine, triethylamine, diethylamine,ethylamine, tributylamine, pyridine, N,N-dimethylaniline,N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine,dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine andN,N′-dibenzylethylenediamine, ethylenediamine, ethanolamine,diethanolamine, piperidine, piperazine, and the like.

Doses and Administration

Compositions used in the invention may be formulated as pharmaceuticalcompositions or formulations for oral, aerosol, parenteral,subcutaneous, intravenous, intramuscular, intraarterial, intrathecal,interperitoneal, nasal, rectal, topical, or vaginal administration. Theterm “parenteral” as used herein includes subcutaneous, intravenous,intramuscular, intra-articular, intra-synovial, intrasternal,intrathecal, intrahepatic, intralesional and intracranial injection orinfusion techniques. Preferably, the compositions are administeredorally, intraperitoneally or intravenously.

The pharmaceutical compositions and formulations may includepharmaceutically acceptable carriers. The term “pharmaceuticallyacceptable carrier,” as used herein, means a non-toxic, inert solid,semi-solid or liquid filler, diluent, encapsulating material orformulation auxiliary of any type. Some examples of materials which canserve as pharmaceutically acceptable carriers are sugars such as, butnot limited to, lactose, glucose and sucrose; starches such as, but notlimited to, corn starch and potato starch; cellulose and its derivativessuch as, but not limited to, sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin;talc; excipients such as, but not limited to, cocoa butter andsuppository waxes; oils such as, but not limited to, peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols; such as propylene glycol; esters such as, but notlimited to, ethyl oleate and ethyl laurate; agar; buffering agents suchas, but not limited to, magnesium hydroxide and aluminum hydroxide;alginic acid; pyrogen-free water; isotonic saline; Ringer's solution;ethyl alcohol, and phosphate buffer solutions, as well as othernon-toxic compatible lubricants such as, but not limited to, sodiumlauryl sulfate and magnesium stearate, as well as coloring agents,releasing agents, coating agents, sweetening, flavoring and perfumingagents, preservatives and antioxidants can also be present in thecomposition, according to the judgment of the formulator.

Thus, the compounds and their physiologically acceptable salts may beformulated for administration by, for example, solid dosing, eye drop,in a topical oil-based formulation, injection, inhalation (eitherthrough the mouth or the nose), implants, or oral, buccal, parenteral,or rectal administration. Techniques and formulations may generally befound in “Remington's Pharmaceutical Sciences,” (Meade Publishing Co.,Easton, Pa.). Therapeutic compositions must typically be sterile andstable under the conditions of manufacture and storage.

The route by which the disclosed compounds are administered and the formof the composition will dictate the type of carrier to be used. Thecomposition may be in a variety of forms, suitable, for example, forsystemic administration (e.g., oral, rectal, nasal, sublingual, buccal,implants, or parenteral) or topical administration (e.g., dermal,pulmonary, nasal, aural, ocular, liposome delivery systems, oriontophoresis).

Carriers for systemic administration typically include at least one ofdiluents, lubricants, binders, disintegrants, colorants, flavors,sweeteners, antioxidants, preservatives, glidants, solvents, suspendingagents, wetting agents, surfactants, combinations thereof, and others.

Formulations suitable for oral administration can consist of (a) liquidsolutions; (b) capsules, sachets, tablets, lozenges, and troches, eachcontaining a predetermined amount of the active ingredient, as solids orgranules; (c) powders; (d) suspensions in an appropriate liquid; and (e)suitable emulsions. Liquid formulations may include diluents, such aswater and alcohols, for example, ethanol, benzyl alcohol, and thepolyethylene alcohols, either with or without the addition of apharmaceutically acceptable surfactant. Capsule forms can be of theordinary hard- or soft-shelled gelatin type containing, for example,surfactants, lubricants, and inert fillers, such as lactose, sucrose,calcium phosphate, and corn starch. Tablet forms can include one or moreof lactose, sucrose, mannitol, corn starch, potato starch, alginic acid,microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicondioxide, croscarmellose sodium, talc, magnesium stearate, calciumstearate, zinc stearate, stearic acid, and other excipients, colorants,diluents, buffering agents, disintegrating agents, moistening agents,preservatives, flavoring agents, and other pharmacologically compatibleexcipients. Lozenge forms can comprise the therapeutic agent(s) in aflavor, usually sucrose and acacia or tragacanth, as well as pastillescomprising the therapeutic agent(s) in an inert base, such as gelatinand glycerin, or sucrose and acacia, emulsions, gels, and the likecontaining, in addition to, such excipients as are known in the art.

The therapeutic agent(s), alone or in combination with other suitablecomponents, can be made into aerosol formulations to be administered viainhalation. These aerosol formulations can be placed into pressurizedacceptable propellants, such as dichlorodifluoromethane, propane,nitrogen, and the like. They also may be formulated as pharmaceuticalsfor non-pressured preparations, such as in a nebulizer or an atomizer.Such spray formulations also may be used to spray mucosa.

Formulations suitable for parenteral administration include aqueous andnon-aqueous, isotonic sterile injection solutions, which can containanti-oxidants, buffers, bacteriostats, and solutes that render theformulation isotonic with the blood of the intended recipient, andaqueous and nonaqueous sterile suspensions that can include suspendingagents, solubilizers, thickening agents, stabilizers, and preservatives.The therapeutic agent can be administered in a physiologicallyacceptable diluent in a pharmaceutical carrier, such as a sterile liquidor mixture of liquids, including water, saline, aqueous dextrose andrelated sugar solutions, an alcohol, such as ethanol or hexadecylalcohol, a glycol, such as propylene glycol or polyethylene glycol,dimethylsulfoxide, glycerol, ketals such as2,2-dimethyl-1,3-di-oxolane-4-methanol, ethers, poly(ethyleneglycol)400, oils, fatty acids, fatty acid esters or glycerides, or acetylatedfatty acid glycerides with or without the addition of a pharmaceuticallyacceptable surfactant, such as a soap or a detergent, suspending agent,such as pectin, carbomers, methylcellulose,hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifyingagents and other pharmaceutical adjuvants.

Oils, which can be used in parenteral formulations include any blandfixed oil, e.g., petroleum, animal, vegetable, synthetic oils, orsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. Specificexamples of oils include peanut, soybean, sesame, cottonseed, corn,olive, petrolatum, and mineral. Suitable fatty acids for use inparenteral formulations include oleic acid, stearic acid, and isostearicacid. Ethyl oleate and isopropyl myristate are examples of suitablefatty acid esters. These oil solutions or suspensions may also contain along-chain alcohol diluent or dispersant, such as carboxymethylcellulose or similar dispersing agents that are commonly used in theformulation of pharmaceutically acceptable dosage forms includingemulsions and suspensions.

Suitable soaps for use in parenteral formulations include fatty alkalimetal, ammonium, and triethanolamine salts, and suitable detergentsinclude (a) cationic detergents such as, for example, dimethyl dialkylammonium halides, and alkyl pyridinium halides, (b) anionic detergentssuch as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin,ether, and mono glyceride sulfates, and sulfosuccinates, (c) nonionicdetergents such as, for example, fatty amine oxides, fatty acidalkanolamides, and polyoxyethylenepolypropylene copolymers, (d)amphoteric detergents such as, for example, alkyl-P-aminopropionates,and 2-alkyl-imidazoline quaternary ammonium salts, and (e) mixturesthereof.

The parenteral formulations will typically contain from about 0.5% toabout 25% by weight of the therapeutic agent(s) in solution.Preservatives and buffers may be used. In order to minimize or eliminateirritation at the site of injection, such compositions may contain oneor more nonionic surfactants having a hydrophile-lipophile balance (HLB)of from about 12 to about 17. The quantity of surfactant in suchformulations will typically range from about 5% to about 15% by weight.Suitable surfactants include polyethylene glycol sorbitan fatty acidesters, such as sorbitan monooleate and the high molecular weightadducts of ethylene oxide with a hydrophobic base, formed by thecondensation of propylene oxide with propylene glycol. The parenteralformulations can be presented in unit-dose or multi-dose sealedcontainers, such as ampoules and vials, and can be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid excipient, for example, water, for injections,immediately prior to use. Extemporaneous injection solutions andsuspensions can be prepared from sterile powders, granules, and tablets.

Additionally, the therapeutic agent(s), or compositions comprisingtherapeutic agent, can be made into suppositories by mixing with avariety of bases, such as emulsifying bases or water-soluble bases.Formulations suitable for vaginal administration can be presented aspessaries, tampons, creams, gels, pastes, foams, or spray formulascontaining, in addition to the active ingredient, such carriers as areknown in the art to be appropriate.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of compounds of this inventioninclude, but are not limited to, mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene, polyoxypropylenecompound, emulsifying wax and water. Alternatively, providedpharmaceutically acceptable compositions can be formulated in a suitablelotion or cream containing the active components suspended or dissolvedin one or more pharmaceutically acceptable carriers. Suitable carriersinclude, but are not limited to, mineral oil, sorbitan monostearate,polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol,benzyl alcohol and water.

For purposes of all of the inventive methods, the administered amount ordose of the therapeutic agent(s) should be sufficient to effect atherapeutic response in the subject or animal over a reasonable timeframe. For example, the dose of the therapeutic agent(s) should besufficient to prevent or inhibit metastasis in a period of from about 2hours or longer, e.g., 12 to 24 or more hours, from the time ofadministration. In certain embodiments, the time period could be evenlonger. The dose will be determined by the efficacy of the particulartherapeutic or agent and the condition of the animal (e.g., human), aswell as the body weight of the animal (e.g., human) to be treated. Manyassays for determining an administered dose are known in the art. Forpurposes of the invention, an assay, which comprises comparing theextent to which the metastasis of a cancer cell is inhibited uponadministration of a given dose of a therapeutic agent to a mammal amonga set of mammals of which is each given a different dose of thetherapeutic agent could be used to determine a starting dose to beadministered to a mammal. The extent to which the metastasis of a cancercell is inhibited or to which the tumor growth is inhibited uponadministration of a certain dose can be assayed by methods known in theart.

The dose of the therapeutic agent also will be determined by theexistence, nature and extent of any adverse side effects that mightaccompany the administration of a particular therapeutic agent.Typically, the attending physician will decide the dosage of thetherapeutic agent with which to treat each individual patient, takinginto consideration a variety of factors, such as age, body weight,general health, diet, sex, therapeutic agent to be administered, routeof administration, and the severity of the condition being treated. Byway of example and not intending to limit the present invention, thedose of the therapeutic agent can be about 0.001 to about 1000 mg/kgbody weight of the subject being treated/day, from about 0.01 to about10 mg/kg body weight/day, about 0.01 mg to about 1 mg/kg bodyweight/day, about 1 to about 100 mg/kg body weight/day, about 10 toabout 90 mg/kg body weight/day, about 20 to about 80 mg/kg bodyweight/day, about 30 to about 70 mg/kg body weight/day, about 40 toabout 60 mg/kg body weight/day, about 50 mg/kg body weight/day, about100 to about 400 mg/kg body weight/day, about 200 to about 300 mg/kgbody weight/day, or about 250 mg/kg body weight/day.

EXAMPLES

Association Between Thromboxane A2 Receptor Gain of Function andMetastasis

Data from a Phenome Wide Association Study (PheWAS) show that a singlenucleotide polymorphism (SNP) in the gene TBXA2R causes again-of-function variant (T399A) in the thromboxane A2 receptor stronglyassociating with increased tumor metastasis (N=32; P=0.003) (Table 1).FIG. 7 shows the primary tumor sites that resulted in the metastaticspread in patients having the gain of function variant T399A. In someembodiments, the invention provides a method of inhibiting solid tumormetastasis comprising administering to a subject with the T399A variant,an amount of a thromboxane A2 receptor antagonist effective to inhibitmetastasis of a solid tumor in the subject.

TABLE 1 Odds Case Total Condition Code P-value Ratio Carriers CasesControls Secondary malignant neoplasm 198 3.8E−03 1.95 32 3568 20663Secondary malignancy of respiratory 198.2 6.5E−03 2.42 13 1190 20663organs Secondary malignancy of lymph nodes 198.1 7.0E−03 2.2  17 169820663 Secondary malignant neoplasm of 198.3 1.2E−02 2.85  7  520 20663digestive systems Secondary malignancy of brain/spine 198.5 2.1E−02 2.68 7  587 20663

Ifetroban Decreases Both Macro- and Micrometastasis in the 4T1 MouseModel of Metastasis

1×10⁵ 4T1-RFP-luciferase cells were implanted into the left inguinalmammary fatpads of 6-week old, virgin WT Balb/C female mice using thefollowing protocol. Mammary tumor cells (4T1-RFP-luciferase cells) arecollected by trypsinization, counted, and resuspended in 50 microlitersof serum-free media+50 microliters of growth factor-reduced Matrigel,for a total volume of 100 microliters. The cells are loaded into asyringe with a 26-g needle. The cells are injected into the inguinalmammary gland, which lies directly under (and attached to) the skin. Theentire injection procedure takes about 10 seconds per mouse. Mice weretreated by orogastric gavage with ifetroban (50 mg/kg daily) or vehiclecontrol when average tumor volume reached 200 mm 3, using 8 mice in theifetroban treatment group, 4 mice in the control group, and 4% sucrosein water for the vehicle. Tumors, lungs and plasma were harvestedapproximately 3 weeks later. Lungs were assessed by whole mounthematoxylin staining to enumerate surface metastases (macro metastases)(FIG. 1 ). Lungs were sectioned and stained with hematoxylin and eosinto confirm that hematoxylin-stained nodules were metastatic lesions, andto enumerate microscopic metastases (FIG. 2 ).

Ifetroban Decreases Macrometastasis in the 4T1 Mouse Model of Metastasis(Second Experiment)

1×10⁵ 4T1-RFP-luciferase cells were implanted into the left inguinalmammary fatpads of 6-week old, virgin WT Balb/C female mice as above.Mice were treated by orogastric gavage with ifetroban (50 mg/kg daily)or vehicle control when average tumor volume reached mm 3 as above.Additionally a third group was treated with aspirin (12 mg/kg daily).Lungs were harvested approximately 4 weeks later. Lungs were assessed bywhole mount hematoxylin staining to enumerate surface metastases (macrometastases) (FIG. 3 ).

Ifetroban decreases metastases in MDA-MB-231 mice. Mice were randomizedinto groups receiving treatment by orogastric gavage with ifetroban (50mg/kg daily) or vehicle, using 10 mice in both the ifetroban and controlgroups and 4% sucrose in water for the vehicle. Mice were treated 48 hprior to hematogenous delivery of MDA-MB-231 cells (1×10⁵ cells) by tailvein injection, and ifetroban treatment was continued for 3 weeksthereafter, at which point lungs were collected from each mouse, andlung metastatic lesions visible to the naked eye were counted (FIG. 4 ).

Ifetroban Treatment does not Affect Primary Tumor Volume, MouseBodyweight or the Survival of 4T1 Cells in Culture

Cell culture experiments (in triplicate, repeated three times) performedusing 4T1 cells treated for 96 hours with ifetroban, revealed noifetroban-mediated changes in 4T1 cell number (automated cell counting),or in the number of apoptotic 4T1 cells (Annexin V staining) (FIG. 5A).For cell culture experiments, vehicle is PBS.

1×10⁵ 4T1-RFP-luciferase cells were implanted into the left inguinalmammary fat pads of 6-week old, virgin WT Balb/C female mice. Mice weretreated by orogastric gavage with ifetroban (50 mg/kg daily; N=8) orvehicle control (N=4) when average tumor volume reached 200 mm 3.Ifetroban treatment (50 mg/kg daily) for 3 weeks did not affect primarytumor volume (FIG. 5B). Mouse total body weight was not affected byIfetroban treatment (50 mg/kg daily) for 3 weeks (FIG. 5C).

Ifetroban decreased hematogenous metastasis of breast, pancreatic andlung cancer cells. MDA-MB-231 (breast), MiaPaca2 (pancreas) and A549(lung) cancer cells were delivered by tail vein injection to nu/nu micepre-treated 24 hours by orogastric gavage with ifetroban (50 mg/kg; N=10per group) or vehicle (N=10). Mice continued daily treatment for anadditional 21 days. On day 21, lungs were harvested and assessed formetastases. N=9, P value, Student's t-test. The results are shown inFIG. 8 .

Ifetroban in Treating Patients with Malignant Solid Tumors at High Riskof Metastatic Recurrence (Prophetic Example).

SUMMARY

A pilot trial studies the side effects of ifetroban in treating patientswith malignant solid tumors that are at high risk of coming back aftertreatment and spreading throughout the body. Platelets are a type ofblood cells that help with clotting. Cancer cells stick to platelets andride on them to get to different parts of the body. Drugs, such asifetroban, may help these platelets become less “sticky,” and reduce thechance of cancer cells spreading to other places in the body.

Detailed Study Description

PRIMARY OBJECTIVES: To assess the safety and feasibility of ifetrobansodium (ifetroban) administration in patients with malignant solidtumors at high risk of metastatic recurrence, after completion of allplanned (neo)adjuvant locoregional and systemic therapies.

SECONDARY OBJECTIVES: To assess rate of metastatic recurrence aftercompletion of ifetroban in patients with malignant solid tumors.

EXPLORATORY OBJECTIVES: To quantify pharmacodynamic markers of ifetrobaneffects.

OUTLINE: 60 patients are randomized to 1 of 2 groups. GROUP 1(IFETROBAN): Patients receive 250 mg ifetroban sodium capsule orally(PO) once daily (QD). Courses repeat every 28 days for 12 months in theabsence of disease progression or unacceptable toxicity. GROUP 2(PLACEBO): Patients receive a 250 mg placebo capsule PO QD. Coursesrepeat every 28 days for 12 months in the absence of disease progressionor unacceptable toxicity. After completion of study treatment, patientsare followed up at 30 days, then up to 12 months.

Outcome Measures. Primary Outcome Measures are: (1) Incidence of adverseevents (Time Frame up to 30 days after completing treatment); (2)Adherence to treatment (participants will be provided a pill diary torecord when they take their medication; study staff will collect thepill diary from participants at their clinic visits) (Time Frame up to12 months); and (3) Summarized change of FACT-G score (scale=0 to 4)(Time Frame up to 12 months). Secondary Outcome Measures are (1)Percentage of patients within metastatic recurrence (within each cohort)(Time Frame at 12 months); and (2) Event-free survival (within eachcohort) (Time Frame up to 12 months).

Eligibility Criteria

-   -   All adults aged 18 years or older.

Inclusion Criteria:

-   -   Signed and dated written informed consent.    -   Eastern Cooperative Oncology Group (ECOG) performance status 0,        1 or 2.    -   One of the following current diagnoses:        -   Stage IIa to III triple negative breast cancer (TNBC).        -   Stage I to II pancreatic adenocarcinoma.        -   Lung Cancer: Stage IIa to III non-small cell lung cancer            (NSCLC) or limited stage small cell lung cancer (SCLC).        -   Stage IIa to III esophageal or gastroesophageal (GE)            junction cancers (squamous cell carcinoma [SCCA] or            adenocarcinoma).        -   Stage IIa to III stomach cancer.    -   Patients must have completed all standard locoregional and        systemic therapy for their cancer.    -   Administration of an investigational agent prior to enrollment        needs to be completed at least 30 days prior to enrollment.    -   Patients must have recovered (≤grade 1 toxicities) from effects        of local (surgery, radiation) or systemic treatments.    -   Platelet count≥100,000 per mL of blood.    -   Hemoglobin ≥9/g/dL (may have been transfused).    -   Serum creatinine ≤1.5× upper limit of normal (ULN) or estimated        creatinine clearance >mL/min as calculated using the        Cockcroft-Gault (CG) equation.    -   Total serum bilirubin ≤1.5 times upper limit of normal (ULN).    -   Aspartate aminotransferase (AST/serum glutamic oxaloacetic        transaminase [SGOT]) and alanine aminotransferase (ALT/serum        glutamate-pyruvate transaminase [SGPT])≤2.5× ULN.    -   International normalized ratio (INR) below upper limit of normal        (ULN).    -   Female patients of childbearing potential and non-sterile males        must agree to use at least two methods of acceptable        contraception from 15 days prior to first trial treatment        administration until at least 5 months after study participant's        final dose of study drug. Females of childbearing potential are        defined as those who are not surgically sterile or        post-menopausal (i.e. patient has not had a bilateral tubal        ligation, a bilateral oophorectomy, or a complete hysterectomy;        or has not been amenorrheic for 12 months without an alternative        medical cause). Post-menopausal status in females under 55 years        of age should be confirmed with a serum follicle-stimulating        hormone (FSH) level within laboratory reference range for        postmenopausal women. Non-sterile males are those who have not        had a vasectomy with documentation of the absence of sperm in        the ejaculate.    -   Patients unable to read/write in English are eligible to        participate in the overall study but will not participate in the        Patient-Reported Outcome questionnaires throughout the trial.    -   Re-enrollment of a subject that has discontinued the study as a        pre-treatment screen failure (i.e. a consented patient who did        not receive study drugs) is permitted. If reenrolled, the        subject must be re-consented. Only the screening procedures        performed outside of protocol-specified timing must be repeated.

Exclusion Criteria:

-   -   Clinical evidence of residual or distant disease after        completion of standard treatment.    -   Current use of anti-platelet drugs (acetylsalicylic acid [ASA],        nonsteroidal anti-inflammatory drugs [NSAIDs], clopidogrel,        argatroban, etc.) or anticoagulants (warfarin, heparin products,        etc.).    -   Active malignancy within 5 years prior to current diagnosis        except for in situ disease or cancer with very high curability        rate (i.e. testicular cancer, etc.).    -   Uncontrolled co-morbid serious systemic illnesses that in the        opinion of the investigator could compromise therapeutic safety.    -   No concurrent anticancer therapy. Required washout from prior        therapy:        -   Chemotherapy: 21 days.        -   Major surgery: 14 days (provided wound healing is adequate).        -   Radiation: 7 days.        -   Investigational/Biologic Therapy: 30 days.    -   Current symptomatic congestive heart failure (New York Heart        Association >class II), unstable cardiac arrhythmia requiring        therapy (e.g. medication or pacemaker), unstable angina (e.g.        new, worsening or persistent chest discomfort), or uncontrolled        hypertension (systolic >160 mmHg or diastolic >100 mmHg). Or any        of the following occurring within 6 months (180 days) prior to        first dose of study drugs: Myocardial infarction,        coronary/peripheral artery bypass graft, cerebrovascular        accident or transient ischemic attack. (Use of antihypertensive        medication to control blood pressure is allowed.)    -   Ongoing peptic ulcer disease requiring treatment. History of        gastrointestinal bleed. Severe gastro-esophageal reflux disease        requiring treatment.    -   History of bleeding diathesis.    -   Planned elective major surgical intervention while taking        ifetroban.    -   Pregnant or breastfeeding females.    -   Prisoners or subjects who are involuntarily incarcerated.    -   Known psychiatric condition, social circumstance, or other        medical condition reasonably judged by the patient's study        physician to unacceptably increase the risk of study        participation; or to prohibit the understanding or rendering of        informed consent or anticipated compliance with scheduled        visits, treatment schedule, laboratory tests and other study        requirements.

While several embodiments of the present invention have been describedand illustrated herein, those of ordinary skill in the art will readilyenvision a variety of other means and/or structures for performing thefunctions and/or obtaining the results and/or one or more of theadvantages described herein, and each of such variations and/ormodifications is deemed to be within the scope of the present invention.More generally, those skilled in the art will readily appreciate thatall parameters, dimensions, materials, and configurations describedherein are meant to be exemplary and that the actual parameters,dimensions, materials, and/or configurations will depend upon thespecific application or applications for which the teachings of thepresent invention is/are used. Those skilled in the art will recognize,or be able to ascertain using no more than routine experimentation, manyequivalents to the specific embodiments of the invention describedherein. It is, therefore, to be understood that the foregoingembodiments are presented by way of example only and that, within thescope of the appended claims and equivalents thereto, the invention maybe practiced otherwise than as specifically described and claimed. Thepresent invention is directed to each individual feature, system,article, material, kit, and/or method described herein. In addition, anycombination of two or more such features, systems, articles, materials,kits, and/or methods, if such features, systems, articles, materials,kits, and/or methods are not mutually inconsistent, is included withinthe scope of the present invention.

For reasons of completeness, various aspects of the invention are setout in the following numbered clauses:

Clause 1. A method of inhibiting solid tumor metastasis comprisingadministering to a subject in need thereof, an amount of a thromboxaneA2 receptor antagonist, or a pharmaceutically acceptable salt orcomposition thereof, effective to inhibit metastasis of a solid tumor inthe subject.

Clause 2. The method of clause 1, wherein the amount of the thromboxaneA2 receptor antagonist, or the pharmaceutically acceptable salt orcomposition thereof, is effective to inhibit the formation ofcirculating tumor cell clusters.

Clause 3. The method of clause 1 or 2, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit movement ofcirculating tumor cell clusters.

Clause 4. The method of any of clauses 1-3, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit the aggregation ofcirculating tumor cell clusters with platelets.

Clause 5. The method of any of clauses 1-4, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit integrin- and/orselectin-mediated cell survival signaling.

Clause 6. The method of any of clauses 1-5, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to reduce the rate ofmetastatic recurrence.

Clause 7. The method of any of clauses 1-6, wherein the thromboxanereceptor antagonist is ifetroban, GR32191, SQ29548, sulotroban,daltroban, linotroban, ramatroban, seratrodast, terutroban, Z-235,LCB-2853, SQ28668, ICI 192605, AH23848, 0N03708, CPI-211, or pinaneTXA2.

Clause 8. The method of clause 7, wherein the thromboxane receptorantagonist is ifetroban.

Clause 9. The method of any of clauses 1-6, wherein the thromboxanereceptor antagonist is a compound of formula (I),

-   -   including all stereoisomers thereof, wherein    -   m is 1, 2, or 3;    -   n is 0, 1, 2, 3, or 4;

-   -   Z is —(CH₂)₂—, —CH═CH—, or    -   Y is O, a single bond, or —CH═CH—;    -   R is CO₂H, —CO₂C₁₋₆alkyl, CH₂OH, —CONHSO₂R³, —CONHR^(3a), or        —CH₂-tetrazol-5-yl;    -   R³ is C₁₋₆alkyl, 6- to 10-membered aryl, or -L¹-(6- to        10-membered aryl);    -   R^(3a) is C₁₋₆alkyl, 6- to 10-membered aryl, or -L¹-(6- to        10-membered aryl);    -   X is O, S, or NH;    -   R¹ is hydrogen, C₁₋₈alkyl, C₂₋₈alkenyl, C₂₋₈alkynyl,        —(CH₂)t-C(O)—NHR^(1a), or —(CH₂)t-NH—C(O)R^(1a);    -   G¹ is a 6- to 10-membered aryl, a C₃₋₁₂cycloalkyl, a 5- to        12-membered heteroaryl, or a 4- to 12-membered heterocyclyl;    -   L¹ is C₁₋₆alkylene;    -   t is 1 to 12;    -   R^(1a) is C₁₋₆alkyl, C₃₋₁₂cycloalkyl, 42-C₃₋₁₂cycloalkyl, or a        6- to 10-membered aryl;    -   R² is hydrogen, C₁₋₆alkyl, 6- to 10-membered aryl, or -L¹-(6- to        10-membered aryl);    -   or R¹ and R² together with the nitrogen to which they attach        form a 4- to 8-membered heterocyclic ring;    -   wherein each aryl is independently and optionally substituted        with 1 or 2 substituents independently selected from the group        consisting of halogen, cyano, C₁₋₆alkyl, C1-6haloalkyl, OH,        —OC₁₋₆alkyl, —OC1-6haloalkyl, —SC₁₋₆alkyl, —S(O)C₁₋₆alkyl,        —S(O)₂C₁₋₆alkyl, —OC1-6alkylene-phenyl, —S-phenyl, —S(O)-phenyl,        and —S(O)₂-phenyl;    -   wherein each cycloalkyl is independently and optionally        substituted with 1-4 substituents independently selected from        the group consisting of halogen, C₁₋₆alkyl, C1-6haloalkyl, OH,        and —OC₁₋₆alkyl.

Clause 10. The method of clause 9, wherein the compound of formula (I)has formula (I-h)

Clause 11. The method of clause 9 or 10, wherein R is CO₂H, or an alkalimetal salt thereof.

Clause 12. The method of any of clauses 9-11, wherein m is 1, n is 2, R²is hydrogen, and R¹ is C₁₋₈alkyl.

Clause 13. The method of any of clauses 9-12, wherein X is O.

Clause 14. The method of any of clauses 1-13, wherein the subject has aprimary tumor of a cancer selected from the group consisting of lungcancer, non-small cell lung cancer, breast cancer, ovarian cancer,prostate cancer, testicular cancer, pancreatic cancer, melanoma,sarcoma, cervical cancer, endometrial cancer, liver cancer, uterinecancer, kidney cancer, gastroesophageal cancer, colon cancer, bladdercancer, mouth cancer, and throat cancer.

Clause 15. The method of any of clauses 1-14, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit metastasis of thesolid tumor in the subject without inhibiting the growth or developmentof the solid tumor.

Clause 16. The method of any of clauses 1-15, further comprisingadministering at least one chemotherapeutic agent chosen from analkylating agent, an anti-metabolite, an anti-tumor antibiotic, ananti-cytoskeletal agent, a topoisomerase inhibitor, an antihormonalagent, a targeted therapeutic agent, immunotherapy, and combinationsthereof.

Clause 17. The method of any of clauses 1-16, wherein the thromboxane A2receptor antagonist, or the pharmaceutically acceptable salt orcomposition thereof, is administered after a chemotherapy treatmentregimen.

Clause 18. The method of any of the foregoing clauses, wherein thesubject has a T399A gain of function mutation of the thromboxane A2receptor.

Clause 19. A thromboxane A2 receptor antagonist, or a pharmaceuticallyacceptable salt or composition thereof, for use in the treatment orinhibition of solid tumor metastasis in a subject.

Clause 20. A thromboxane A2 receptor antagonist, or a pharmaceuticallyacceptable salt or composition thereof, for use in a method of treatingor inhibiting solid tumor metastasis, wherein the method comprisesadministering the thromboxane A2 receptor antagonist to a subject inneed thereof.

Clause 21. Use of a thromboxane A2 receptor antagonist, or apharmaceutically acceptable salt or composition thereof, for thepreparation of a medicament for the treatment or inhibition of solidtumor metastasis in a subject.

We claim:
 1. A method of inhibiting solid tumor metastasis comprisingadministering to a subject in need thereof, an amount of a thromboxaneA2 receptor antagonist, or a pharmaceutically acceptable salt orcomposition thereof, effective to inhibit metastasis of a solid tumor inthe subject.
 2. The method of claim 1, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit the formation ofcirculating tumor cell clusters.
 3. The method of claim 1, wherein theamount of the thromboxane A2 receptor antagonist, or thepharmaceutically acceptable salt or composition thereof, is effective toinhibit movement of circulating tumor cell clusters.
 4. The method ofclaim 1, wherein the amount of the thromboxane A2 receptor antagonist,or the pharmaceutically acceptable salt or composition thereof, iseffective to inhibit the aggregation of circulating tumor cell clusterswith platelets.
 5. The method of claim 1, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit integrin- and/orselectin-mediated cell survival signaling.
 6. The method of claim 1,wherein the amount of the thromboxane A2 receptor antagonist, or thepharmaceutically acceptable salt or composition thereof, is effective toreduce the rate of metastatic recurrence.
 7. The method of claim 1,wherein the thromboxane receptor antagonist is ifetroban, GR32191,SQ29548, sulotroban, daltroban, linotroban, ramatroban, seratrodast,terutroban, Z-235, LCB-2853, SQ28668, ICI 192605, AH23848, ON03708,CPI-211, or pinane TXA₂.
 8. The method of claim 7, wherein thethromboxane receptor antagonist is ifetroban.
 9. The method of claim 1,wherein the thromboxane receptor antagonist is a compound of formula(I),

including all stereoisomers thereof, wherein m is 1, 2, or 3; n is 0, 1,2, 3, or 4; Z is —(CH₂)₂—, —CH═CH—, or

Y is O, a single bond, or —CH═CH—; R is CO₂H, —CO₂C₁₋₆alkyl, CH₂OH,—CONHSO₂R³, —CONHR^(3a), or —CH₂—tetrazol-5-yl; R³ is C₁-6alkyl, 6- to10-membered aryl, or -L¹-(6- to 10-membered aryl); R^(3a) is C₁₋₆alkyl,6- to 10-membered aryl, or -L¹-(6- to 10-membered aryl); X is O, S, orNH; R¹ is hydrogen, C₁₋₈alkyl, C₂₋₈alkenyl, C₂₋₈alkynyl, G¹, -L¹-G¹,—(CH₂)_(t)—C(O)—NHR^(1a), or —(CH₂)_(t)NH—C(O)R^(1a); G¹ is a 6- to10-membered aryl, a C₃₋₁₂cycloalkyl, a 5- to 12-membered heteroaryl, ora 4- to 12-membered heterocyclyl; L¹ is C₁₋₆alkylene; t is 1 to 12;R^(1a) is C₁₋₆alkyl, C₃₋₁₂cycloalkyl, -L¹-C₃₋₁₂cycloalkyl, or a 6- to10-membered aryl; R² is hydrogen, C₁₋₆alkyl, 6- to 10-membered aryl, or-L¹-(6- to 10-membered aryl); or R¹ and R² together with the nitrogen towhich they attach form a 4- to 8-membered heterocyclic ring; whereineach aryl is independently and optionally substituted with 1 or 2substituents independently selected from the group consisting ofhalogen, cyano, C₁₋₆alkyl, C₁₋₆haloalkyl, OH, —OC₁₋₆alkyl,—OC₁₋₆haloalkyl, —SC₁₋₆alkyl, —S(O)C₁₋₆alkyl, —S(O)₂C₁₋₆alkyl, —OC₁₋₆alkylene-phenyl, —S—phenyl, —S(O)-phenyl, and —S(O)₂-phenyl; whereineach cycloalkyl is independently and optionally substituted with 1-4substituents independently selected from the group consisting ofhalogen, C₁₋₆alkyl, C₁₋₆haloalkyl, OH, and —OC₁₋₆alkyl.
 10. The methodof claim 9, wherein the compound of formula (I) has formula (I-h)


11. The method of claim 10, wherein R is CO₂H, or an alkali metal saltthereof.
 12. The method of claim 11, wherein m is 1, n is 2, R² ishydrogen, and R¹ is C₁-8alkyl.
 13. The method of claim 12, wherein X isO.
 14. The method of claim 8, wherein the subject has a primary tumor ofa cancer selected from the group consisting of lung cancer, non-smallcell lung cancer, breast cancer, ovarian cancer, prostate cancer,testicular cancer, pancreatic cancer, melanoma, sarcoma, cervicalcancer, endometrial cancer, liver cancer, uterine cancer, kidney cancer,gastroesophageal cancer, colon cancer, bladder cancer, mouth cancer, andthroat cancer.
 15. The method of claim 1, wherein the amount of thethromboxane A2 receptor antagonist, or the pharmaceutically acceptablesalt or composition thereof, is effective to inhibit metastasis of thesolid tumor in the subject without inhibiting the growth or developmentof the solid tumor.
 16. The method of claim 1, further comprisingadministering at least one chemotherapeutic agent chosen from analkylating agent, an anti-metabolite, an anti-tumor antibiotic, ananti-cytoskeletal agent, a topoisomerase inhibitor, an antihormonalagent, a targeted therapeutic agent, immunotherapy, and combinationsthereof.
 17. The method of claim 1, wherein the thromboxane A2 receptorantagonist, or the pharmaceutically acceptable salt or compositionthereof, is administered after a chemotherapy treatment regimen.
 18. Themethod of claim 1, wherein the subject has a T399A gain of functionmutation of the thromboxane A2 receptor. 19-21. (canceled)